中华鳖诱导型一氧化氮合酶(iNOS)基因的表达分析  

Expression analysis of the inducible nitric oxide synthase(iNOS)gene of the Pelodiscus sinensis

作　　者：夏美文 吕孙建 李逸群 齐天鹏 徐海圣[1,3] XIA Meiwen;LYU Sunjian;LI Yiqun;QI Tianpeng;XU Haisheng(College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;Institute of Hydrobiology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;South Taihu Lake Modern Agricultural Science and Technology Extension Center of Huzhou,Huzhou 313000,Zhejiang,China)

机构地区：[1]浙江大学动物科学学院,杭州310058 [2]浙江省农业科学院/水生生物研究所,杭州310021 [3]湖州市南太湖现代农业科技推广中心,浙江湖州313000

出　　处：《淡水渔业》2024年第6期64-70,共7页Freshwater Fisheries

基　　金：湖州市科技计划项目(2023ZD2037);湖州市农业科技创新项目(2019HN02)。

摘　　要：一氧化氮(NO)参与生物体内多种重要的生理病理活动,可增强机体的免疫功能,而诱导型一氧化氮合酶(iNOS)是一氧化氮合成过程中的关键限速酶。为了探明iNOS在中华鳖(Pelodiscus sinensis)先天免疫中的作用,研究了iNOS基因在中华鳖组织的分布及外源刺激物对其基因和蛋白表达的影响。荧光定量PCR结果显示,iNOS基因在中华鳖的心脏、肝脏、肾脏、脾脏、肌肉、小肠和大肠中均有表达,其中在大肠中表达量最高,其次是小肠、肾脏和脾脏。在受脂多糖(lipopolysaccharides,LPS)刺激后,肝、脾、肾、肌肉和小肠iNOS mRNA的表达显著升高。免疫组化实验结果显示LPS刺激后12 h、24 h iNOS蛋白在小肠、脾脏表达水平显著高于生理盐水对照组。研究结果表明iNOS可能参与了中华鳖的机体免疫反应。Considered one of China’s commercially important aquatic species of China,the Chinese softshell turtle(Pelodiscus sinensis)contributes a high portion of the total aquatic products sold in the country.P.sinensis is directly exposed to the aquatic environment,and because the water is very nutrient-rich,it is vulnerable to infection by opportunistic pathogens.Nitric oxide(NO)participates in various important physiological and pathological activities in organisms.It can enhance the immune function of the body,and inducible nitric oxide synthase(iNOS)is a key rate-limiting enzyme in the process of nitric oxide synthesis.In some organisms,it has been demonstrated that treatment with lipopolysaccharides(LPS),a gram-negative bacteria cell product,can lead to a pronounced increase in iNOS expression.The study investigated the distribution of iNOS genes in P.sinensis tissues and the effects of exogenous stimuli on their gene and protein expression to investigate the role of iNOS in the innate immunity of P.sinensis.Quantitative real-time PCR(qRT-PCR)was used to examine the distribution of iNOS in various turtle tissues,including the muscle,spleen,liver,heart,kidney,and small and large intestines.The results of qPCR showed that the iNOS gene was expressed in the heart,liver,kidney,spleen,muscle,and small and large intestines.And the highest expression level was in the large intestine,followed by the small intestine,kidney,and spleen.P.sinensis were equally distributed into two groups.The challenge group was chosen to receive an injection of LPS,while the control group was given of saline injections.Five turtles from each group were dissected to collect the liver,small intestine,spleen,kidney,and muscle at various intervals(0,3,6,12,24,and 48 hours).After LPS stimulation,iNOS mRNA expression was significantly elevated in the liver,spleen,kidney,muscle,and small intestine.The results of IHC showed that the expression levels of iNOS protein in the small intestine and spleen were significantly higher than that in the saline-t

关 键 词：中华鳖(Pelodiscus sinensis) 诱导型一氧化氮合酶 脂多糖 

分 类 号：S947.1[农业科学—水产养殖]